The cell cycle of the planctomycete Gemmata obscuriglobus with respect to cell compartmentalization.

Lee KC, Webb RI, Fuerst JA - BMC Cell Biol. (2009)

Bottom Line:
Fluorescence microscopy of DAPI-stained cells of G. obscuriglobus suggested that translocation of the nucleoid and formation of the bud did not occur at the same time.The new bud nucleoid is initially naked and not surrounded by membrane, but eventually acquires a complete nucleoid envelope consisting of two closely apposed membranes as occurs in the mother cell.The membranes of the new nucleoid envelope surrounding the bud nucleoid are derived from intracytoplasmic membranes of both the mother cell and the bud.

Affiliation: School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia. s4010907@student.uq.edu.au

ABSTRACT

Background: Gemmata obscuriglobus is a distinctive member of the divergent phylum Planctomycetes, all known members of which are peptidoglycan-less bacteria with a shared compartmentalized cell structure and divide by a budding process. G. obscuriglobus in addition shares the unique feature that its nucleoid DNA is surrounded by an envelope consisting of two membranes forming an analogous structure to the membrane-bounded nucleoid of eukaryotes and therefore G. obscuriglobus forms a special model for cell biology. Draft genome data for G. obscuriglobus as well as complete genome sequences available so far for other planctomycetes indicate that the key bacterial cell division protein FtsZ is not present in these planctomycetes, so the cell division process in planctomycetes is of special comparative interest. The membrane-bounded nature of the nucleoid in G. obscuriglobus also suggests that special mechanisms for the distribution of this nuclear body to the bud and for distribution of chromosomal DNA might exist during division. It was therefore of interest to examine the cell division cycle in G. obscuriglobus and the process of nucleoid distribution and nuclear body formation during division in this planctomycete bacterium via light and electron microscopy.

Results: Using phase contrast and fluorescence light microscopy, and transmission electron microscopy, the cell division cycle of G. obscuriglobus was determined. During the budding process, the bud was formed and developed in size from one point of the mother cell perimeter until separation. The matured daughter cell acted as a new mother cell and started its own budding cycle while the mother cell can itself initiate budding repeatedly. Fluorescence microscopy of DAPI-stained cells of G. obscuriglobus suggested that translocation of the nucleoid and formation of the bud did not occur at the same time. Confocal laser scanning light microscopy applied to cells stained for membranes as well as DNA confirmed the behaviour of the nucleoid and nucleoid envelope during cell division. Electron microscopy of cryosubstituted cells confirmed deductions from light microscopy concerning nucleoid presence in relation to the stage of budding, and showed that the nucleoid was observed to occur in both mother and bud cells only at later budding stages. It further suggested that nucleoid envelope formed only after the nucleoid was translocated into the bud, since envelopes only appeared in more mature buds, while naked nucleoids occurred in smaller buds. Nucleoid envelope appeared to originate from the intracytoplasmic membranes (ICM) of both mother cell and bud. There was always a connecting passage between mother cell and bud during the budding process until separation of the two cells. The division cycle of the nucleated planctomycete G. obscuriglobus appears to be a complex process in which chromosomal DNA is transported to the daughter cell bud after initial formation of the bud, and this can be performed repeatedly by a single mother cell.

Conclusion: The division cycle of the nucleated planctomycete G. obscuriglobus is a complex process in which chromosomal nucleoid DNA is transported to the daughter cell bud after initial formation of a bud without nucleoid. The new bud nucleoid is initially naked and not surrounded by membrane, but eventually acquires a complete nucleoid envelope consisting of two closely apposed membranes as occurs in the mother cell. The membranes of the new nucleoid envelope surrounding the bud nucleoid are derived from intracytoplasmic membranes of both the mother cell and the bud. The cell division of G. obscuriglobus displays some unique features not known in cells of either prokaryotes or eukaryotes.

Figure 6: Gemmata obscuriglobus cell budding cycle. In (A) and (B), a bud initiates from one point of the mother cell M1 enclosing a nucleoid (blue) surrounded by a double-membrane nucleoid envelope (grey). The mother cell ICM (dark purple) is continuous with that of the bud (also dark purple). In (C), the naked nucleoid is translocated into the bud at some early budding stage. In (D), the bud nucleoid is surrounded by two membranes where an inner membrane (light purple) continuous with mother cell ICM, and an outer membrane (also light purple) shows continuity with the bud ICM. In (E), the bud nucleoid is completely surrounded by the two closely apposed membranes where membrane fusion and pinching off has resulted in a double-membrane nucleoid envelope completely separated from ICM membranes. (E) is the end-point of a possible model mechanism where the bud reached similar cell size as M1. (F) shows the separation of the mother cell and the matured bud. In (G) and (H), the mother cell M1 can initiate the next budding cycle after a 2–4 hour lag (lag 1) while the matured bud M2 can begin its first budding cycle after a 3–5.5 hour lag (lag2).

Mentions:
Little is known about the cell division cycle of the compartmentalized planctomycetes, especially how membrane-bounded compartments such as those enclosing the nucleoid are transferred into the daughter cells. Results of phase contrast and fluorescence light microscopy combined with electron microscopy of thin-sectioned cells prepared by high pressure freezing/cryosubstitution can be used to derive a model for the cell cycle of Gemmata obscuriglobus. The G. obscuriglobus cell cycle is summarized in Fig. 6. In this model a mother cell forms a small bud with a narrow neck relative to mother cell diameter, and this bud gradually enlarges until it is similar in size to the mother cell, a stage which then lasts for a time considerably longer than other stages of cell division. Both the mother cell and the finally released bud are capable of further cell division – the bud can only start this after a lag period which is much longer than the lag observed for the second mother cell budding. There appears to be a distinct reproductive pole, since division seems to occur repeatedly at the same pole. A new bud is formed at the same pole position of the mother cell where the previous bud was formed, matured and separated from the mother cell.

Figure 6: Gemmata obscuriglobus cell budding cycle. In (A) and (B), a bud initiates from one point of the mother cell M1 enclosing a nucleoid (blue) surrounded by a double-membrane nucleoid envelope (grey). The mother cell ICM (dark purple) is continuous with that of the bud (also dark purple). In (C), the naked nucleoid is translocated into the bud at some early budding stage. In (D), the bud nucleoid is surrounded by two membranes where an inner membrane (light purple) continuous with mother cell ICM, and an outer membrane (also light purple) shows continuity with the bud ICM. In (E), the bud nucleoid is completely surrounded by the two closely apposed membranes where membrane fusion and pinching off has resulted in a double-membrane nucleoid envelope completely separated from ICM membranes. (E) is the end-point of a possible model mechanism where the bud reached similar cell size as M1. (F) shows the separation of the mother cell and the matured bud. In (G) and (H), the mother cell M1 can initiate the next budding cycle after a 2–4 hour lag (lag 1) while the matured bud M2 can begin its first budding cycle after a 3–5.5 hour lag (lag2).

Mentions:
Little is known about the cell division cycle of the compartmentalized planctomycetes, especially how membrane-bounded compartments such as those enclosing the nucleoid are transferred into the daughter cells. Results of phase contrast and fluorescence light microscopy combined with electron microscopy of thin-sectioned cells prepared by high pressure freezing/cryosubstitution can be used to derive a model for the cell cycle of Gemmata obscuriglobus. The G. obscuriglobus cell cycle is summarized in Fig. 6. In this model a mother cell forms a small bud with a narrow neck relative to mother cell diameter, and this bud gradually enlarges until it is similar in size to the mother cell, a stage which then lasts for a time considerably longer than other stages of cell division. Both the mother cell and the finally released bud are capable of further cell division – the bud can only start this after a lag period which is much longer than the lag observed for the second mother cell budding. There appears to be a distinct reproductive pole, since division seems to occur repeatedly at the same pole. A new bud is formed at the same pole position of the mother cell where the previous bud was formed, matured and separated from the mother cell.

Bottom Line:
Fluorescence microscopy of DAPI-stained cells of G. obscuriglobus suggested that translocation of the nucleoid and formation of the bud did not occur at the same time.The new bud nucleoid is initially naked and not surrounded by membrane, but eventually acquires a complete nucleoid envelope consisting of two closely apposed membranes as occurs in the mother cell.The membranes of the new nucleoid envelope surrounding the bud nucleoid are derived from intracytoplasmic membranes of both the mother cell and the bud.

Affiliation:
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia. s4010907@student.uq.edu.au

ABSTRACT

Background: Gemmata obscuriglobus is a distinctive member of the divergent phylum Planctomycetes, all known members of which are peptidoglycan-less bacteria with a shared compartmentalized cell structure and divide by a budding process. G. obscuriglobus in addition shares the unique feature that its nucleoid DNA is surrounded by an envelope consisting of two membranes forming an analogous structure to the membrane-bounded nucleoid of eukaryotes and therefore G. obscuriglobus forms a special model for cell biology. Draft genome data for G. obscuriglobus as well as complete genome sequences available so far for other planctomycetes indicate that the key bacterial cell division protein FtsZ is not present in these planctomycetes, so the cell division process in planctomycetes is of special comparative interest. The membrane-bounded nature of the nucleoid in G. obscuriglobus also suggests that special mechanisms for the distribution of this nuclear body to the bud and for distribution of chromosomal DNA might exist during division. It was therefore of interest to examine the cell division cycle in G. obscuriglobus and the process of nucleoid distribution and nuclear body formation during division in this planctomycete bacterium via light and electron microscopy.

Results: Using phase contrast and fluorescence light microscopy, and transmission electron microscopy, the cell division cycle of G. obscuriglobus was determined. During the budding process, the bud was formed and developed in size from one point of the mother cell perimeter until separation. The matured daughter cell acted as a new mother cell and started its own budding cycle while the mother cell can itself initiate budding repeatedly. Fluorescence microscopy of DAPI-stained cells of G. obscuriglobus suggested that translocation of the nucleoid and formation of the bud did not occur at the same time. Confocal laser scanning light microscopy applied to cells stained for membranes as well as DNA confirmed the behaviour of the nucleoid and nucleoid envelope during cell division. Electron microscopy of cryosubstituted cells confirmed deductions from light microscopy concerning nucleoid presence in relation to the stage of budding, and showed that the nucleoid was observed to occur in both mother and bud cells only at later budding stages. It further suggested that nucleoid envelope formed only after the nucleoid was translocated into the bud, since envelopes only appeared in more mature buds, while naked nucleoids occurred in smaller buds. Nucleoid envelope appeared to originate from the intracytoplasmic membranes (ICM) of both mother cell and bud. There was always a connecting passage between mother cell and bud during the budding process until separation of the two cells. The division cycle of the nucleated planctomycete G. obscuriglobus appears to be a complex process in which chromosomal DNA is transported to the daughter cell bud after initial formation of the bud, and this can be performed repeatedly by a single mother cell.

Conclusion: The division cycle of the nucleated planctomycete G. obscuriglobus is a complex process in which chromosomal nucleoid DNA is transported to the daughter cell bud after initial formation of a bud without nucleoid. The new bud nucleoid is initially naked and not surrounded by membrane, but eventually acquires a complete nucleoid envelope consisting of two closely apposed membranes as occurs in the mother cell. The membranes of the new nucleoid envelope surrounding the bud nucleoid are derived from intracytoplasmic membranes of both the mother cell and the bud. The cell division of G. obscuriglobus displays some unique features not known in cells of either prokaryotes or eukaryotes.